Kito M, Ohishi N, Yagi K
Institute of Applied Biochemistry, Mitake, Gifu, Japan.
Biochem Int. 1991 May;24(1):131-5.
Highly sensitive assay method of L-gulono-gamma-lactone oxidase (GLO) was constructed. In this method, L-ascorbic acid formed in the enzymatic reaction was converted to its bis(dinitrophenyl)hydrazone derivative, and the amount of the latter was determined by high-performance liquid chromatography. Twenty picomoles of ascorbic acid was detected, which makes this method 25 times more sensitive than the previously used dipyridyl one. By the present method, a minute activity of GLO in liver microsomes prepared from rats of the Osteogenic Disorder Shionogi strain (ODS-od/od) could be measured.
构建了L-古洛糖酸-γ-内酯氧化酶（GLO）的高灵敏度检测方法。在该方法中，酶促反应中形成的L-抗坏血酸被转化为其双（二硝基苯基）腙衍生物，后者的量通过高效液相色谱法测定。检测到20皮摩尔的抗坏血酸，这使得该方法的灵敏度比先前使用的联吡啶法高25倍。通过本方法，可以测量从成骨障碍柴田株（ODS-od/od）大鼠制备的肝微粒体中微量的GLO活性。
